P
US11873316B2ActiveUtilityPatentIndex 83

Compositions and methods for phosphoramidite and oligonucleotide synthesis

Assignee: WAVE LIFE SCIENCES LTDPriority: Nov 23, 2016Filed: Nov 22, 2017Granted: Jan 16, 2024
Est. expiryNov 23, 2036(~10.4 yrs left)· nominal 20-yr term from priority
Inventors:BUTLER DAVID CHARLES DONNELLKANDASAMY PACHAMUTHUMARAPPAN SUBRAMANIANPAIK IK-HYEONKUMARASAMY JAYAKANTHANBOMMINENI GOPAL REDDY
C07H 1/06C07H 19/10C07H 19/207C07H 21/00C07F 9/65844C07H 1/00C07H 19/16
83
PatentIndex Score
10
Cited by
265
References
18
Claims

Abstract

The present disclosure, among other things, provides technologies for oligonucleotide synthesis. In some embodiments, the present disclosure provides phosphoramidites and methods for synthesis thereof. In some embodiments, provided methods provides higher yields and/or purities. In some embodiments, provided methods remove byproducts without contact with an aqueous solution.

Claims

exact text as granted — not AI-modified
The invention claimed is: 
     
       1. A method for preparing a phosphoramidite, comprising steps of:
 a) providing a preparation comprising a phosphoramidite product and one or more reaction byproducts of the synthesis of the phosphoramidite; and 
 b) performing a filtration step on the preparation to provide a filtrate, wherein the filtrate contains a reduced level of the one or more reaction byproducts relative to the product as compared to that level present in the preparation prior to the filtration step, 
 wherein: 
 after the phosphoramidite product is formed, no solution comprising the phosphoramidite product is contacted with water of more than 100 mole equivalents of the phosphoramidite; and 
 the phosphoramidite has the structure of formula I: 
 
       
         
           
           
               
               
           
         
         wherein: 
         BA is 
       
       
         
           
           
               
               
           
         
          wherein R is hydrogen or optionally substituted C 1-6  aliphatic; 
         SU is 
       
       
         
           
           
               
               
           
         
         R 5 s is R′ or —OR′; 
         wherein: 
         (1) R 2s  is —F or —OCH 2 CH 2 OMe, and 
       
       
         
           
           
               
               
           
         
          or 
         (2) R 2s  is —F, —H, or —OCH 2 CH 2 OMe, and 
       
       
         
           
           
               
               
           
         
         each R′ is independently —R, —C(O)R, —CO 2 R, or —SO 2 R, or:
 two or more R′ are taken together with their intervening atoms to form an optionally substituted monocyclic, bicyclic or polycyclic, saturated, partially unsaturated, or aryl 3-30 membered ring having, in addition to the intervening atoms, 0-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon; and 
 
         each R is independently hydrogen, or an optionally substituted group selected from C 1-30  aliphatic, C 1-30  heteroaliphatic having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, C 6-30  aryl, a 5-30 membered heteroaryl ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon, and a 3-30 membered heterocyclic ring having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus and silicon. 
       
     
     
       2. The method of  claim 1 , wherein the preparation comprises a solution and one or more solids. 
     
     
       3. The method of  claim 2 , wherein the solution comprises the phosphoramidite product. 
     
     
       4. The method of  claim 3 , wherein the one or more solids comprise one or more reaction byproducts. 
     
     
       5. The method of  claim 4 , wherein R 5S  is DMTrO-. 
     
     
       6. The method of  claim 5 , wherein R 2s  is —H. 
     
     
       7. The method of  claim 4 , comprising purifying the phosphoramidite product using silica gel column chromatography, wherein the silica gel is treated with an anhydrous acetonitrile. 
     
     
       8. The method of  claim 5 , wherein R 2s  is —F. 
     
     
       9. The method of  claim 8 , wherein 
       
         
           
           
               
               
           
         
       
     
     
       10. The method of  claim 8 , wherein 
       
         
           
           
               
               
           
         
       
     
     
       11. The method of  claim 5 , wherein R 2s  is —OCH 2 CH 2 OMe. 
     
     
       12. The method of  claim 11 , wherein 
       
         
           
           
               
               
           
         
       
     
     
       13. The method of  claim 11 , wherein BA is 
       
         
           
           
               
               
           
         
       
     
     
       14. The method of  claim 4 , wherein BA is 
       
         
           
           
               
               
           
         
       
       in which R is C 1-6  alkyl. 
     
     
       15. The method of  claim 4 , wherein BA is 
       
         
           
           
               
               
           
         
       
       in which R is isopropyl. 
     
     
       16. The method of  claim 1 , wherein the prepared phosphoramidite has a purity of 90% or more. 
     
     
       17. The method of  claim 1 , wherein the prepared phosphoramidite has a purity of 95% or more. 
     
     
       18. The method of  claim 1 , wherein the prepared phosphoramidite has a purity of 96% or more.

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